ARTICLE TOOLS

Abstract

Fluorescence imaging is the most powerful technique available for observing spatial and temporal dynamics of ions and molecules in the single living cells. However, almost all the molecular events, including those involving second messengers and protein phosphorylation, are analyzed by conventional destructive methods, such as radio immunoassay and immunoblotting of cell lysates prepared from millions of cells. Here, we describe fluorescent reporters for protein phosphorylation and lipid second messengers that we recently developed, and show how their spatial and temporal dynamics can be monitored in single living cells. The present fluorescent reporters should be an indispensable tool not only for understanding the complex mechanism of the signal transduction in single cells and organisms but also for high-throughput screening of pharmaceutical candidates that regulate the cellular extent of protein phosphorylation and lipid second messengers.